牙鲆感染迟钝爱德华氏菌的miRNA转录组分析  

Analysis of miRNA transcriptome in Paralichthys olivaceus infected with Edwardsiella tarda

作　　者：山亚男 郑津辉[1,2] 高虹[1,2] SHAN Yanan;ZHENG Jinhui;GAO Hong(College of Life Sciences,Tianjin Normal University,Tianjin 300387,China;Tianjin Key Laboratory of Animal and Plant Resistance,Tianjin Normal University,Tianjin 300387,China)

机构地区：[1]天津师范大学生命科学学院,天津300387 [2]天津师范大学天津市动植物抗性重点实验室,天津300387

出　　处：《天津师范大学学报（自然科学版）》2022年第2期37-45,共9页Journal of Tianjin Normal University：Natural Science Edition

基　　金：天津市自然科学基金重点资助项目(19JCZDJC34300,14JCZDJC34200).

摘　　要：为了探究牙鲆感染迟钝爱德华氏菌过程中miRNA的分子调控机制,以牙鲆头肾组织为研究材料,分析其感染迟钝爱德华氏菌后miRNA的差异表达情况.利用高通量测序平台(Illumina HiSeq)分别对感染迟钝爱德华氏菌3 h、6 h以及未感染的牙鲆头肾组织的RNA进行转录组测序,并对数据进行生物信息学分析.将测序结果与斑马鱼miRBase数据库进行比对,共鉴定出牙鲆成熟miRNA 537个.将感染组与对照组数据进行比对,得到12个感染组(3 h)差异显著表达的miRNA(2个显著上调,10个显著下调);32个感染组(6 h)差异显著表达的miRNA(20个显著上调,12个显著下调).对感染组差异显著表达的miRNA的靶基因进行GO seq分析,结果显示,感染组(3 h)的富集通路有2793条(P<0.05),包括2021个生物过程、315个细胞组分和457个分子功能;感染组(6 h)的富集通路有3048条(P<0.05),包括2235个生物过程、342个细胞组分和471个分子功能.KEGG分析结果显示,感染组(3 h)中获得71个富集通路(P<0.05),包括31个有机系统、17个环境信息处理、6个细胞过程、13个人类疾病和4个代谢通路;感染组(6 h)中获得69个富集通路(P<0.05),包括33个有机系统、18个环境信息处理、6个细胞过程、10个人类疾病和2个代谢通路.选取6个差异表达miRNA进行实时荧光定量PCR验证,结果显示,荧光定量结果与转录组测序结果趋势一致.总的来看,在牙鲆感染迟钝爱德华氏菌过程中,差异表达miRNA的靶基因参与的信号通路大部分跟生物过程以及细胞组分中的膜成分有关,生物过程中候选靶基因富集程度最高的GO条目是生物调节,富集程度较高的通路大多跟免疫调节以及分裂分化等生物学过程有关.To investigate the molecular regulation mechanism of miRNA in Paralichthys olivaceus infected with Edwardsiella tarda,the differential expression of miRNA in P.olivaceus after infection by E.tarda was analyzed.Transcriptome sequencing was performed using high-throughput sequencing platform(Illumina HiSeq)for RNA from head kidney tissue of P.olivaceus infected by E.tarda at 3 h,6 h and non-infected individual,and the data were analyzed by bioinformatics.A total of 537 mature miRNAs were identified by comparing the sequencing results with miRBase database of Danio rerio.Comparing the data of the infection group and the control group,12 miRNAs were differentially expressed in infection group(3 h)(2 up-regulated and 10 down-regulated);32 miRNAs were differentially expressed in infection group(6 h)(20 up-regulated and 12 down-regu-lated).GO seq analysis results of target genes of differentially expressed miRNA in the infection group showed that there were 2793 miRNA enrichment pathways in infection group(3 h)(P<0.05),including 2021 biological processes(BP),315 cellular components(CC)and 457 molecular functions(MF);There were 3048 enrichment pathways in infection group(6 h)(P<0.05),including 2235 BP,342 CC and 471 MF.KEGG analysis showed that 71 pathways were enriched in infection group(3 h)(P<0.05),including 31 organic systems,17 environmental information processing,6 cellular processes,13 human diseases and 4 metabolic pathways;69 pathways were enriched in infection group(6 h)(P<0.05),including 33 organic sys tems,18 environmental information processing,6 cellular processes,10 human diseases and 2 metabolic pathways.Six differ-entially expressed miRNAs were selected for real-time fluorescence quantitative PCR verification.The results showed that the trend of fluorescence quantitative results was consistent with transcriptome sequencing results.In general,in the process of E.tarda infection of P.olivaceus,most of the signal pathways involved in the target genes differentially expressing miRNA are related to biological

关 键 词：迟钝爱德华氏菌 牙鲆 MIRNA 转录组 高通量 差异表达miRNA 

分 类 号：S943[农业科学—水产养殖]